Differentiation of cGMP-dependent and -independent nitric oxide effects on platelet apoptosis and reactive oxygen species production using platelets lacking soluble guanylyl cyclase

Natalia Rukoyatkina; Ulrich Walter; Andreas Friebe; Stepan Gambaryan

doi:10.1160/TH11-05-0319

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2011; 106(05): 922-933
DOI: 10.1160/TH11-05-0319

Platelets and Blood Cells

Schattauer GmbH

Differentiation of cGMP-dependent and -independent nitric oxide effects on platelet apoptosis and reactive oxygen species production using platelets lacking soluble guanylyl cyclase

Authors

Natalia Rukoyatkina

¹Institute of Clinical Biochemistry and Pathobiochemistry, University of Würzburg, Würzburg, Germany

²Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, St. Petersburg, Russia
Ulrich Walter

¹Institute of Clinical Biochemistry and Pathobiochemistry, University of Würzburg, Würzburg, Germany
Andreas Friebe

³Institute of Physiology, University of Würzburg, Würzburg, Germany
Stepan Gambaryan

¹Institute of Clinical Biochemistry and Pathobiochemistry, University of Würzburg, Würzburg, Germany

²Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, St. Petersburg, Russia

Abstract

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Summary

Platelet activation is an irreversible process resulting in platelet apoptosis and necrosis, and circulating platelets contain many components of the apoptotic machinery. Cyclic guanosine monophosphate (cGMP) generated by nitric oxide (NO) activated soluble guanylyl cyclase (sGC) plays a crucial role in preventing platelet activation. However, in addition to activation of sGC, cGMP-independent NO effects in platelets have been described. To differentiate between cGMP-dependent and -independent NO effects on platelet apoptosis and reactive oxygen species (ROS) production, we generated platelet-specific sGC-deficient mice (PS-GCKO). Platelet apoptosis was induced by a combination of thrombin/convulxin (Thr/Cvx) and assessed by phosphatidylserine (PS) surface exposure, and loss of the mitochondrial membrane potential. NO-induced inhibition of PS externalisation was mediated only by cGMP-dependent mechanisms. Inhibition of the mitochondrial membrane potential decrease at low NO concentration was also cGMP-dependent but became cGMP-independent at high NO concentrations. In contrast, inhibition of ROS formation at any NO concentration was mediated by cGMP-independent mechanisms, very likely due to direct radical scavenging. NO inhibits platelet apoptosis by cGMP-dependent mechanisms and ROS production by cGMP-independent mechanisms. The PS-GCKO mouse model is an important tool for the differentiation of cGMP-dependent and -independent NO effects on platelets.

Keywords

Apoptosis - cGMP - NO - PKG - platelet - sGC

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References

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